Wheat (Triticum aestivum L.) is one of the most important staple crops globally. Doubled haploid technology enables rapid development of pure lines and has been extended from maize to several other crop species. A key step in DH breeding is the identification of haploids from diploids, which requires accurate and convenient phenotypic markers. In this study, we generated two wheat haploid inducers carrying different markers by a one-step strategy. One harbored a dual fluorescent marker system consisting of eGFP and TagRFP, the other carried a RUBY reporter. Both markers enabled near 100% accuracy of haploid identification at the immature embryo, mature embryo, and germinating seedling stages. Moreover, both lines consistently exhibited a high and stable haploid induction rate (~20%). This study not only provides efficient wheat haploid inducers but also establishes a convenient pipeline for developing haploid induction systems in other crop species.

Root hairs are fast growing, ephemeral tubular extensions of the root epidermis that aid nutrient and water uptake. The aim of the present study was to identify QTL for root hair length (RHL) using 227 F₈ recombinant inbred lines (RILs) derived from a cross of Zhou 8425B (Z8425B) and Chinese Spring (CS), and to develop convenient molecular markers for marker-assisted breeding in wheat. Analysis of variance of root hair length showed significant differences (P < 0.01) among RILs. The genetic map for QTL analysis consisted of 3389 unique SNP markers. Using composite interval mapping, four major QTL (LOD > 2.5) for RHL were identified on chromosomes 1B (2), 2D and 6D and four putative QTL (2 ≤ LOD ≤ 2.5) were detected on chromosomes 1A, 3A, 6B, and 7B, explaining 3.32%–6.52% of the phenotypic variance. The positive alleles for increased RHL of QTL on chromosomes 2D, 6B and 6D (QRhl.cau-2D, qRhl.cau-6B, and QRhl.cau-6D) were contributed by Z8425B, and CS contributed positive QTL alleles on chromosomes 1A (qRhl.cau-1A), 1B (QRhl.cau-1B.1 and QRhl.cau-1B.2), 3A (qRhl.cau-3A) and 7B (qRhl.cau-7B). STARP markers were developed for QRhl.cau-1B.1, QRhl.cau-2D, QRhl.cau-6D, and qRhl.cau-7B. Haplotype and association analysis indicated that the positive allele of QRhl.cau-6D had been strongly selected in Chinese wheat breeding programs. Collectively, the identified QTL for root hair length are likely to be useful for marker-assisted selection.

Hexaploid triticale (×Triticosecale, AABBRR) is an important forage crop and a promising energy plant. Transferring D-genome chromosomes or segments from common wheat (Triticum aestivum) into hexaploid triticale is attractive in improving its economically important traits. Here, a hexaploid triticale 6D(6A) substitution line Lin 456 derived from the cross between the octoploid triticale line H400 and the hexaploid wheat Lin 56 was identified and analyzed by genomic in situ hybridization (GISH), fluorescence in situ hybridization (FISH), and molecular markers. The GISH analysis showed that Lin 456 is a hexaploid triticale with 14 rye (Secale cereale) chromosomes and 28 wheat chromosomes, whereas non-denaturing fluorescence in situ hybridization (ND-FISH) and molecular marker analysis revealed that it is a 6D(6A) substitution line. In contrast to previous studies, the signal of Oligo-pSc119.2 was observed at the distal end of 6DL in Lin 456. The wheat chromosome 6D was associated with increased grain weight and decreased spikelet number using the genotypic data combined with the phenotypes of the F₂ population in the three environments. The thousand-grain weight and grain width in the substitution individuals were significantly higher than those in the non-substitution individuals in the F₂ population across the three environments. We propose that the hexaploid triticale 6D(6A) substitution line Lin 456 can be a valuable and promising donor stock for genetic improvement during triticale breeding.